The majority of allelopathic studies on invasive plants have focused primarily on their leaf-mediated allelopathy, with relatively little attention paid to their root-mediated allelopathy, especially co-allelopathy mediated by both leaves and roots. It is conceivable that the diversified composition of acid rain may influence the allelopathy of invasive plants. This study aimed to evaluate the leaf and root-mediated co-allelopathy of the invasive plant Solidago canadensis L. under acid rain with different nitrogen-sulfur ratios (N/S) on Lactuca sativa L. via a hydroponic incubation. The root-mediated allelopathy of S. canadensis was found to be more pronounced than the leaf-mediated allelopathy of S. canadensis with nitric acid at pH 4.5, but the leaf-mediated allelopathy of S. canadensis was observed to be more pronounced than the root-mediated allelopathy of S. canadensis with sulfuric-rich acid at pH 4.5. The leaf and root-mediated co-allelopathy of S. canadensis was more pronounced than that of either part alone with sulfuric acid at pH 5.6 and nitric acid at pH 4.5, but not with nitric-rich acid at pH 4.5 and sulfuric-rich acid at pH 4.5. Sulfuric acid and sulfuric-rich acid with stronger acidity intensified the leaf-mediated allelopathy of S. canadensis. Nitric acid and nitric-rich acid attenuated the leaf-mediated allelopathy of S. canadensis, and most types of acid rain (especially nitric acid and nitric-rich acid) also attenuated the root-mediated allelopathy of S. canadensis and the leaf and root-mediated co-allelopathy of S. canadensis. Sulfuric acid and sulfuric-rich acid produced a more pronounced effect than nitric acid and nitric-rich acid. Hence, the N/S ratio of acid rain influenced the allelopathy of S. canadensis under acid rain with multiple N/S ratios.